N-(1,2,3,4-tetrahydronaphthalen-1-yl)-4-phenyl-1-piperazinealkylamide derivatives, and therapeutic use thereof as 5-HT7 receptor ligands

ABSTRACT

A series of N-(1,2,3,4-tetrahydronaphthalen-1-yl)-4-aryl-1-piperazinealkylamides was prepared and their affinity for serotonin 5-HT 7 , 5-HT 1A , and 5-HT 2A  receptors was measured using in vitro binding assays. In relation to 5-HT 7  receptor affinity, receptor binding studies indicated that: (i) the optimal alkyl chain length was five methylenes; (ii) an unsubstituted 1,2,3,4-tetrahydronaphthalenyl nucleus was selected for further substitutions; and (iii) the substitution pattern of the aryl ring linked to the piperazine ring played a significant role. Several compound with high affinity for 5-HT 7  receptors were identified. Among them, 4-(2-methoxyphenyl)-N-(1,2,3,4-tetrahydronaphthalen-1-yl)-1-piperazinehexanamide (28), 4-(2-acetylphenyl)-N-(1,2,3,4-tetrahydronaphthalen-1-yl)-1-piperazinehexanamide (34), 4-(2-methylthiophenyl)-N-(1,2,3,4-tetrahydronaphthalen-1-yl)-1-piperazinehexanamide (44), 4-(2-hydroxyphenyl)-N-(1,2,3,4-tetrahydronaphthalen-1-yl)-1-piperazinehexanamide (46), 4-(2-methylphenyl)-N-(1,2,3,4-tetrahydronaphthalen-1-yl)-1-piperazinehexanamide (49) were assayed for the 5-HT 7  receptor mediated relaxation of substance P-induced guinea-pig ileum contraction. Compounds 28, 44, and 49 behaved as full agonists, compound 34 as a partial agonist, whereas derivative 46 acted as an antagonist.

FIELD OF THE INVENTION

The present invention relates to a novel family of 5-HT₇ Receptorligands, being derivatives ofN-(1,2,3,4-tetrahydronaphthalen-1-yl)-4-(2-substituted-phenyl)-1-piperazinealkylamide,and to their therapeutic use in the treatment of all those statessuitable to be relieved by 5-HT₇ receptor agonists or antagonists.

BACKGROUND OF THE INVENTION

The neurotransmitter serotonin (5-hydroxytryptamine, 5-HT) has an arrayof pharmacological and physiological roles within the central nervoussystem (CNS) and in the periphery, mediated by its interactions with atotal of 14 structurally and pharmacologically distinct receptorsubtypes. These receptors have been assigned to one of seven families,5-HT₁₋₇. The 5-HT₇ receptor (5-HT₇R) is the most recent addition to the5-HT receptor family, and was cloned for the first time in 1993 from ratand mouse. Since then, it has been cloned from other species such ashuman, guinea-pig, and pig. The 5-HT₇R was shown to be positivelycoupled to adenylyl cyclase via Gs proteins. It displays a low degree ofhomology (40%) with other Gs-coupled 5-HT receptors. Four differentisoforms have been found, namely 5-HT_(7a), 5-HT_(7b), 5-HT_(7c),5-HT_(7d). Only two isoforms (5-HT_(7a) and 5-HT_(7b)) are present inboth rat and human, whereas the 5-HT_(7c) receptor is found exclusivelyin rat, while the 5-HT_(7d) is found only in human. Each of the isoformsappears to form a functionally active receptor with the 5-HT_(7a), beingthe most abundant (80%) in both rat and human brain. There appear to beno pharmacological differences among the four isoforms. Highconcentrations of the 5-HT₇R have been detected by in situ hybridizationand 5-HT₇-like immunoreactivity in the hypothalamus, entorhinal cortex,septal areas, substantia nigra, amygdala, rapes nuclei and thetrigeminal nucleus. In addition, moderate levels of 5-HT₇-likeimmunoreactivity were found in the thalamus, hippocampus, cingulate andoccipital cortex, caudate, putamen, and suprachiasmatic nucleus (SCN) ofthe rat. This distribution correlates well with distribution of mRNAencoding 5-HT₇R protein. In fact, the 5-HT₇R mRNA has been detected inthalamus, hypothalamus, hippocampus, amygdala, cortex, septum, andsuprachiasmatic nucleus.

The potential of therapeutic effects of 5-HT₇ agents have beenhypothesized on the basis of such anatomical distribution. The linkbetween 5-HT₇Rs and the SCN suggests a potential role in circadianrhythms and sleep disorders. Lovenberg et al. (Lovenberg, T. W., Baron,B. M., de Lecea, L., Miller, J. D., Prosser, R. A., Rea, M. A., Foye, P.E., Racke, M., Slone, A. L., Siegel, B. W., Danielson, P. E., Sutcliffe,J. G., Erlander, M. G. Neuron 1993, 11, 449-458) demonstrated that phaseadvances in circadian neuronal activity of the SCN could be elicitedusing serotonergic ligands that display a pharmacological profileconsistent with that of the 5-HT₇R. Since then, 5-HT₇Rs have been shownto be present in postsynaptic areas in the SCN where serotonergicneurones are proposed to play a key role in modulating circadianactivity. Mullins et al.( Mullins, U. L.; Gianutsos, G.; Eison, A. S.Neuropsychopharmacol. 1999, 21, 352-367.) have supplied supportingevidence that implicates a possible role for 5-HT₇R in depression. Theydemonstrated that antidepressant-induced expression of the immediateearly gene, c-Fos, in the SCN was blocked by ritanserin (ahigh-affinity, but non-selective, 5-HT₇R antagonist), but not by the5-HT_(1A) antagonist pindolol or the 5-HT_(1D) antagonist sumatriptan.This suggests that the effect is mediated through 5-HT₇Rs, although,with such non-selective antagonists, the involvement of other 5-HTreceptors cannot be ruled out.

The involvement of the 5-HT₇R in migraine pathogenesis has been proposedby Terron (Terron, J. A. Eur. J. Pharmacol. 2002, 439, 1-11) because the5-HT₇R-mediated vasodilator mechanism operates in vascular structuresthat have been implicated in migraine, such as the middle cerebral andexternal carotid arteries. Finally, several compounds possessing high5-HT₇R affinity have therapeutic indications as antipsychotic drugs andthis has suggested that 5-HT₇R may mediate therapeutic action of suchcompounds (Roth, B. L.; Craigo, S. C.; Choudhary, M. S.; Uluer, A.;Monsma, F. J. Jr.; Shen, Y.; Meltzer, H. Y.; Sibley, D. R. J. Pharmacol.Exp. Ther. 1994, 268, 1403-1410).

It is therefore clear that the 5-HT₇R may be a valuable drug target.During the last decade considerable research efforts have been directedtowards the identification of selective 5-HT₇R antagonists, (Leopoldo,M. Curr. Med. Chem. 2004, 11, 629-661) allowing the identification ofsome interesting compounds such as SB-258719, SB-269970, SB-656104,DR4004, LY215840, the chemical structures of which are depicted in FIG.1.

However, these promising compounds present several limitations becauseof their low potency (SB-258719), modest selectivity (SB-656104,LY215840), and low metabolic stability (SB-269970, DR4004).

Therefore, the scope of the present invention is that of providing novelselectively-acting 5-HT₇R ligands as useful pharmacological tools orpotential drugs.

It is noteworthy that most 5-HT₇R ligands reported to date act asantagonists, whereas a very limited number of agonists has beenreported.

Of the different chemical classes which bind to 5-HT₇Rs,arylpiperazines, four species of which are depicted and numbered in FIG.2, have received our attention as well as that of other authors.Recently, we have reported structure affinity relationship studies oftwo distinct classes of 5-HT₇R ligands, based on the structure of1-arylpiperazine. Examples of these classes are represented by compounds3 and 4 (Perrone, R., Berardi, F., Colabufo, N. A., Lacivita, E.,Leopoldo, M., Tortorella, V. J. Med. Chem. 2003, 46, 646-649; Leopoldo,M.; Berardi, F.; Colabufo, N. A.; Contino, M.; Lacivita, L.; Perrone,R.; Tortorella, V. J. Pharm. Pharmacol. 2004, 56, 247-255).

In the present study, we screened the 1-(2-methoxyphenyl) piperazinederivatives 5-7, previously prepared in our laboratory as 5-HT_(1A)ligands (Perrone, et al. J. Med. Chem. 1996, 39, 3195-3202.

Perrone, R. et al. J. Med. Chem. 1998, 41, 4903-4909) against the clonedrat 5-HT₇R because they share some structural features with derivatives1 and 2. We found that the compounds 6 and 7 possessed moderateaffinities for 5-HT₇R, as well as for 5-HT_(1A) receptor.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 provides the chemical structures of some prior art compoundsdeveloped as selective 5-HT₇R antagonists.

FIG. 2 provides the chemical structures of four species of prior artarylpiperazines.

FIG. 3 provides a depiction of some of the synthetic pathways utilizedin preparation of compounds of the invention.

BRIEF DISCUSSION OF ASPECTS OF THE INVENTION

We here describe the structural modifications of compound 7, inparticular modification in i) the intermediate alkyl chain length, ii)the presence and the position of the methoxy group on the1,2,3,4-tetrahydronaphthalene nucleus, and iii) the position and thetype of the aromatic substituent linked to the N-1 piperazine ring.

This experimental work has resulted in the identification of a novelfamily of high affinity 5-HT₇ receptor ligands based on theN-(1,2,3,4-tetrahydronaphthalen-1-yl)-4-(2substituted-phenyl)-1-piperazinealkylamide structure.

One of several aspects of the present invention is therefore a family of5-HT₇ receptor ligands having general formula I. A second aspect of theinvention are compositions comprising the same ligands. A third aspectis the treatment of a human or animal subject in need thereof with oneor more such compositions for any pathological state suitable of beingrelieved by these ligands or their pharmaceutically acceptable salts orother common derivatives.

The compounds of the invention are characterized by an extremely highaffinity for the receptor 5-HT₇ and a considerable selectivity over5-HT_(1A) and 5-HT_(2A) receptors.

Due to these affinity characteristics, various ligands of the inventionfind applicability in the treatment of those states suitable to berelieved by 5-HT₇ receptor agonists or antagonists.

DETAILED DESCRIPTION OF EMBODIMENTS OF THE INVENTION

Aspects of embodiments of the invention were published by the presentinventors M. Leopoldo et al. in “Journal of Medicine Chemistry”, Vol.47, No. 26 pages 6616-6624, published on web on 19 Nov. 2004. Thispublication is specifically incorporated by reference by reference forany teaching not provided herein. Also, in reviewing the detaileddisclosure which follows, it should be borne in mind that all patents,patent applications, patent publications, technical publications,scientific publications, and other references referenced herein arehereby incorporated by reference in this application in order to morefully describe the state of the art to which the present inventionpertains.

It also is noted that compounds referred to by number correspond to thedescriptions of the compounds as may be found in the accompanying tablesand in the text.

Starting fromN-(5-methoxy-1,2,3,4-tetrahydronaphthalen-1-yl)-4-(2-methoxyphenyl)-1-piperazinebutanamide(7), we have identified a new class of 5-HT₇R ligands. The structuralmodification introduced on 7 allowed the elucidation of the structuralrequirements for high 5-HT₇R affinity of this class of compounds. Inparticular, all structural modifications introduced on either the1,2,3,4-tetrahydronaphthalenyl nucleus or on the linker between thisparticular group and the N-(2-methoxyphenyl)piperazine moiety influencedonly the 5-HT₇R affinity and not the selectivity over 5-HT_(1A)receptor. In contrast, modifications of the aryl group linked to thepiperazine ring resulted in major changes in 5-HT₇R affinity. Therefore,the 4-aryl-N-(1,2,3,4-tetrahydronaphthalen-1-yl)-1-piperazinehexanamidestructure was identified as a promising framework to obtain highaffinity 5-HT₇R ligands. Among the compounds displaying the highest5-HT₇R affinity, derivatives 28, 34, 44, 46, 49 were submitted to afunctional assay to establish their intrinsic activity. Compounds 28,44, and 49 behaved as full agonists, compound 34 as a partial agonist,whereas derivative 46 acted as an antagonist. Among the compoundspresented here,4-(2-methylthiophenyl)-N-(1,2,3,4-tetrahydronaphthalen-1-yl)-1-piperazinehexanamide(44) was identified as a potent 5-HT₇R full agonist (K_(i)=0.22 nM,EC₅₀=2.56 μM), with selectivity over 5-HT_(1A) and 5-HT_(2A) receptors(200-fold and >1000-fold, respectively).

Preparation of the Compounds of the Invention

Some of the synthetic pathways utilized in preparation of the compoundsof the invention are depicted in Scheme 2, shown in FIG. 3. Acylation ofamines 9a - e with the appropriate ω-haloacyl chloride afforded the keyintermediates 10a-c,e, 11a-e, 12a-e that reacted with the appropriate1-arylpiperazine to give the final compounds 7, 15-22, 24-50. Thissynthetic pathway was not useful to obtain derivative 23 in a pure form,therefore an alternative synthetic route was followed:1-(2-methoxyphenyl)piperazine was reacted with ethyl 4-chorobutanoate togive ester 13. Hydrolysis of the latter gave carboxylic acid 14 thatreacted with amine 9d to give the expected final compound. All targetcompounds were prepared as racemates.

The first modification performed on compound 7 was the optimization ofthe intermediate alkyl chain length. Therefore, we evaluated compounds15 and 16 (Table 1) having a four or five methylene alkyl chain,respectively. 5-HT₇R affinity values indicated that alkyl chainelongation resulted in an increasing in affinity. Secondly, we shiftedthe methoxy group from the 5-position to the 6-, 7-, and 8-position ofthe tetrahydronaphthalenyl ring, because previous studies indicated thatthe position of the methoxy group on the terminal aromatic nucleusinfluenced the 5-HT₇R affinity of compounds 3 and 4 (Perrone R. et al.2003 supra; Leopoldo M. et al 2004 supra) This modification wasperformed on the compounds 7, 15, and 16 that displayed good 5-HT₇Raffinities. Considering each group of isomers (i.e. compounds having thesame alkyl chain length), no significant difference in 5-HT₇R affinitywas observed. Moreover, within each group of homologues (i.e. compoundsbearing the methoxy group in the same position) affinity valuesreplicate the affinity rank already noted for the 5-methoxy substitutedderivatives 7, 15, and 16. Because the position of the methoxy group atthe tetrahydronaphthalenyl ring did not exert a significant role on5-HT₇R affinity of the compounds 7, 15-25, we evaluated theunsubstituted derivatives 26, 27, and 28. This modification improved the5-HT₇R affinity.

The results in Table 1 indicate that the modifications of either the1,2,3,4-tetrahydronaphthalenyl nucleus or of the linker between thisgroup and the N-(2-methoxyphenyl)piperazine moiety of compound 7influenced the 5-HT₇R affinity only and not the selectivity over5-HT_(1A) receptor.

Therefore, we focused on the aromatic ring attached to the piperazinenitrogen, bearing in mind that minimal changes in this part of themolecule might result in major changes in 5-HT₇R affinity as well as in5-HT_(1A) and 5-HT_(2A) receptor affinity, as documented (Perrone R. etal. 2003 supra). Because the derivatives with a five methylene linkerdisplayed the higher 5-HT₇R affinity values, we have further modifiedcompound 28. Initially, based on literature data, we substituted the2-methoxyphenyl group with a bicyclic aromatic system, or a2-acetylphenyl, or a 2-cyanophenyl group. The replacement of the2-methoxyphenyl group with a bicyclic aromatic system (Table 2,compounds 29-32) reduced the 5-HT₇R affinity. In particular, it can benoted that the presence of the benzisoxazolyl group was detrimental for5-HT₇ affinity (compound 29), whereas in previous studies we found thatthis particular replacement resulted in the opposite effect (Perrone R.et al 2003 supra). In contrast, compounds 33 and 34 (Table 3) retainedreasonably good 5-HT₇R affinity, but were unselective over 5-HT_(1A)receptors. Moreover, we prepared compounds 35 and 36 that present anadditional substituent in 4- or 3-position of the aromatic ring, becausethis substitution pattern has been reported to be detrimental for5-HT_(1A) receptor affinity. This modification determined a loss in5-HT₇R affinity and no significant improvement in selectivity over5-HT_(1A) receptors. Additionally, we shifted the substituent from the2-position of compounds 28, 33, and 34 to the 3- and 4 position (Table3, derivative 37-42). Binding data of derivatives 37-42 indicate thataffinity for 5-HT₇R strongly depends on the position of the substituent.In fact, the 3-substituted derivatives 37, 39, and 41 are less potent at5-HT₇R than the 2-substituted isomers 28, 33, and 34. The 4-substitutedderivatives 38, 40, and 42 are nearly devoid of 5-HT₇R affinity. Takentogether, these data confirm that this part of the molecule is quitesensitive to minimal structural changes. Subsequently, we evaluatedanalogues of 28 having a substituent in the 2-position other thanmethoxy as well as the unsubstituted derivative (Table 3, derivatives43-50). For this purpose we selected several substituents with differentelectronic properties. Considering the unsubstituted derivative 50 asreference compound, it can be noted that the cyano, chloro and nitrosubstituents (compounds 33, 47, and 48, respectively) did not change the5-HT₇R affinity. In contrast, carboxamido and methylsulfonylsubstituents (derivatives 43 and 45, respectively) caused a drop in5-HT₇R affinity. Substitution of the 2-position by a methoxy, acetyl,methylthio, hydroxy, or methyl group resulted in high affinity 5-HT₇Rligands (derivatives 28, 34, 44, 46, 49, respectively). These dataindicate that the presence of a substituent in the 2-position modulatethe affinity of this class of compounds for 5-HT₇R. The affinity valuesseem not to be related to electronic, steric, or H-bonding properties ofthese substituents. As a result, clear structure-affinity relationshipsare not evident. Moreover, the 5-HT_(1A) receptor affinities ofcompounds 33-50 parallel the 5-HT₇R affinities, whereas 5-HT_(2A)receptor affinities are negligible. Notably, only compound 44 showedconsiderable selectivity over 5-HT_(1A) and 5-HT_(2A) receptors(200-fold and >1000-fold, respectively).

We tested the structurally related compounds 28, 34, 44, 46, and 49 for5-HT₇ intrinsic activity in an isolated guinea-pig ileum assay (Table4). It has been reported that 5-HT₇ agonists produce a dose-dependentguinea-pig ileum relaxation of substance P-induced contraction (Carter,D.; Champney, M.; Hwang, B.; Eglen, R. M. Eur. J. Pharmacol. 1995, 280,243-250). Compounds 28, 44, and 49 behaved as full agonists, compound 34as a partial agonist, whereas derivative 46 acted as an antagonist.These results indicate that the nature of the 2-substituent is relatedto the intrinsic activity. In particular, the difference in intrinsicactivity between hydroxy derivative 46 and the corresponding methoxyderivative 28 might indicate that the H-bonding donor property of thehydroxy is responsible for the antagonistic property of 46. In contrast,an apolar group seems to promote the activation of 5-HT₇R.

Therapeutic Applications

Previous experimental reports demonstrated the potential role of 5-HT₇receptors in circadian rhythms and sleep disorders (Lovenberg et al.supra).

There is also evidence supporting a possible role for 5-HT₇R indepression. Mullins et al. (supra) demonstrated thatantidepressant-induced expression of the immediate early gene, c-Fos, inthe SCN was blocked by ritanserin, a high-affinity, but non-selective,5-HT₇R antagonist. This suggests that the effect is mediated through5-HT₇Rs, although, with such non-selective antagonists, the involvementof other 5-HT receptors cannot be ruled out.

The involvement of the 5-HT₇R in migraine pathogenesis also has beenproposed in literature Terron (supra) because the 5-HT₇R-mediatedvasodilator mechanism operates in vascular structures that have beenimplicated in migraine, such as the middle cerebral and external carotidarteries. Finally, several compounds possessing high 5-HT₇R affinityhave therapeutic indications as antipsychotic drugs and this hassuggested that 5-HT₇R may mediate therapeutic action of such compounds.

The claimed compounds disclosed here are characterized by an extremelyhigh affinity for the receptor 5-HT₇ with agonistic or antagonisticactivity and a considerable selectivity over 5-HT_(1A) and 5-HT_(2A)receptors. These properties make the compounds of the inventioneffective therapeutic principles for the treatment of all thosedisorders in which the contribution of 5-HT₇ receptors is recognized, inparticular in the treatment of circadian rhythms and sleep disorders andin the treatment of depression, psychotic states and migraine.

Pharmaceutical compositions comprising the ligands of the invention arecompositions suitable for oral or parenteral administering. The activecompounds may be formulated with any pharmaceutically acceptableeccipient in any suitable form such as tablets, capsules, pills,granulates, powder or aqueous, hydroalcolic, oleous solutions or W/O orO/W emulsions or dispersions.

Specific embodiments of the invention are solid compositions for oraluse or liquid solutions for parenteral use comprising the compounds:4-(2-methoxyphenyl)-N-(1,2,3,4-tetrahydronaphthalen-1-yl)-1-piperazinehexanamide(28),4-(2-acetylphenyl)-N-(1,2,3,4-tetrahydronaphthalen-1-yl)-1-piperazinehexanamide(34),4-(2-methylthiophenyl)-N-(1,2,3,4-tetrahydronaphthalen-1-yl)-1-piperazinehexanamide(44),4-(2-hydroxyphenyl)-N-(1,2,3,4-tetrahydronaphthalen-1-yl)-1-piperazinehexanamide(46),4-(2-methylphenyl)-N-(1,2,3,4-tetrahydronaphthalen-1-yl)-1-piperazinehexanamide(49). Among these, particular embodiments of the invention arecompositions comprising the compound (44) that was identified as themost potent 5-HT₇ receptor agonist (K_(i)=0.22 nM, EC₅₀=2.56 μM),endowed with selectivity over 5-HT_(1A) and 5-HT_(2A) receptors(200-fold and >1000-fold, respectively).

EXAMPLES AND ADDITIONAL DISCLOSURE

Experimental Section

The following compounds were synthesized according to publishedprocedures (See M. Leopoldo et al. “Journal of Medicine Chemistry”published on web on 19 Nov. 2004): 1-(2-acetylphenyl)piperazine,1-(3-acetylphenyl)piperazine, 2-bromo(methylsulfonyl)benzene,1-(2-carboxamidophenyl)piperazine,1-(4-chloro-2-methoxyphenyl)piperazine, 1-(2-cyanophenyl)piperazine,1-(3-cyanophenyl)piperazine, 1-(2,5-dimethoxyphenyl)piperazine,5-methoxy-1,2,3,4-tetrahydro-1-naphthalenamine,6-methoxy-1,2,3,4-tetrahydro-1-naphthalenamine,7-methoxy-1,2,3,4-tetrahydro-1-naphthalenamine,8-methoxy-1,2,3,4-tetrahydro-1-naphthalenamine,1-(2-methylthiophenyl)piperazine, 1-(2-nitrophenyl)piperazine,2-(1-piperazinyl)-1H-benzimidazole, 3-(1-piperazinyl)-1,2-benzisoxazole,2-(1-piperazinyl)benzoxazole.

Column chromatography was performed with 1:30 ICN silica gel 60A (63-200μm) as the stationary phase. Melting points were determined in opencapillaries on a Gallenkamp electrothermal apparatus. Elemental analyses(C,H,N) were performed on Eurovector Euro EA 3000 analyzer; theanalytical results were within ±0.4% of the theoretical values for theformula given. ¹H NMR spectra were recorded at 300 MHz on a Bruker AM300 WB spectrometer or on a Varian Mercury-VX spectrometer. All chemicalshift values are reported in ppm (δ). Recording of mass spectra was doneon an HP6890-5973 MSD gas chromatograph/mass spectrometer; onlysignificant. m/z peaks, with their percentage of relative intensity inparentheses, are reported. Compounds 42, 43, and 45 were characterizedby ESI⁺/MS/MS with an Agilent 1100 Series LC-MSD trap System VLworkstation. All spectra were in accordance with the assignedstructures. The purity of new compounds that were essential to theconclusions drawn in the text were determined by HPLC on a Perkin-Elmerseries 200 LC instrument using a Phenomenex Prodigy ODS-3 RP-18 column,(250

4.6 mm, 5 μm particle size) and equipped with a Perkin-Elmer 785A UV/VISdetector setting λ=254 nm. All compounds were eluted withCH₃OH/H₂O/EtN₃, 4:1:0.01, v/v, at a flow rate of 1 mL/min. A standardprocedure was used to transform final compounds into their hydrochlorideor oxalate salts that were recrystallized as detailed in Tables 1-3.

1-[2-(Methylsulfonyl)phenyl]piperazine (8)

A mixture of 2-bromo(methylsulfonyl)benzene (1.10 g, 4.7 mmol) andanhydrous piperazine (2.02 g, 23.5 mmol) was heated at 110° C.overnight. Then, the mixture was cooled and partitioned between 2 N NaOHand CH₂Cl₂. The separated organic layer was dried over Na₂SO₄ andconcentrated under reduced pressure. The crude residue waschromatographed (CHCl₃/CH₃OH, 9:1, as eluent) to give 8 as a whitesemisolid (0.36 g, 34% yield). ¹H NMR: δ 2.58 (s, 1H, NH, D₂Oexchanged), 2.84 (s, 4H, piperazinic), 3.14 (s, 3H, CH₃), 7.10-7.83 (m,4H, aromatic).

General Procedure for Preparation of Alkylating Agents 10a-c,e, 11a-e,12a-e

A cooled solution of amine 9 (4.0 mmol) in CH₂Cl₂ was stirred vigorouslywith 2% aqueous NaOH (9.6 mL, 4.8 mmol) while the appropriateω-haloalkyl chloride (4.8 mmol) in CH₂Cl₂ was added dropwise. The sameNaOH solution was then used to maintain pH at 9, and at constant pH thelayers were separated. The organic phase was washed with 3 N HCl, withH₂O, and then dried over Na₂SO₄ and evaporated under reduced pressure.The crude residue was chromatographed as detailed below to givecompounds 10a-c,e, 11a-e, 12a-e as white semisolids.

N-(5-Methoxy-1,2,3,4-tetrahydronaphthalen-1-yl)-4-chlorobutanamide (10a)

Eluted with CHCl₃/AcOEt, 1:1; 39% yield. ¹H NMR: δ 1.72-1.86, 1.92-2.04(m, 4H, endo CH₂CH₂), 2.10-2.19 (m, 2H, CH₂CH₂CH₂) 2.37 (t, 2H, J=7.2Hz, COCH₂), 2.53-2.79 (m, 2H, benzylic CH₂), 3.63 (t, 2H, J=6.0 Hz,CH₂Cl), 3.82 (s, 3H, CH₃), 5.15-5.20 (m, 1H, CH), 5.75 (br d, 1H, NH),6.72-7.18 (m, 3H, aromatic). GC-MS m/z 283 (M⁺+2, 1), 281 (M⁺, 2), 161(26), 160 (100), 159 (27).

N-(5-Methoxy-1,2,3,4-tetrahydronaphthalen-1-yl)-5-chloropentanamide(11a)

Eluted with CH₂Cl₂; 33% yield. ¹H NMR: δ 1.75-1.85, 1.93-2.01 (m, 8H,CH₂(CH₂)₂CH₂, endo CH₂CH₂), 2.19-2.26 (m, 2H, COCH₂), 2.55-2.73 (m, 2H,benzylic CH₂), 3.52-3.58 (m, 2H, CH₂Cl), 3.81 (s, 3H, CH₃), 5.14-5.19(m, 1H, CH), 5.73 (br d, 1H, NH), 6.71-7.17 (m, 3H, aromatic). GC-MS m/z297 (M⁺+2, 2), 295 (M⁺, 5), 161 (28) 160 (100), 159 (31), 145 (20).

N-(5-Methoxy-1,2,3,4-tetrahydronaphthalen-1-yl)-6-bromohexanamide (12a)

Eluted with CH₂Cl₂; 35% yield. ¹H NMR: δ 1.43-1.53 [m, 2H,(CH₂)₂CH₂(CH₂)₂], 1.61-1.99 [m, 8H, CH₂CH₂Br, COCH₂CH₂, endo CH₂CH₂],2.20 (t, 2H, J=7.4 Hz, COCH₂), 2.53-2.75 (m, 2H, benzylic CH₂), 3.40 (t,2H, J=6.7 Hz, CH₂Br), 3.81 (s, 3H, CH₃), 5.17 (br t, 1H, CH), 5.69 (brd, 1H, NH), 6.74-7.17 (m, 3H, aromatic). GC-MS m/z 355 (M⁺+2, 1), 353(M⁺, 1), 160 (100).

Ethyl 4-[4-(2-methoxyphenyl)piperazin-1-yl]butanoate (13)

A stirred mixture of 1-(2-methoxyphenyl)piperazine (1.50 g, 7.8 mmol),ethyl 4-bromobutanoate (0.9 mL, 6.3 mmol), and K₂CO₃ (0.87 g, 6.3 mmol)in acetonitrile was refluxed overnight. After the mixture was cooled,the mixture was evaporated to dryness and H₂O (20 mL) was added to theresidue. The aqueous phase was extracted with CH₂Cl₂ (2

30 mL). The collected organic layers were dried over Na₂SO₄ andevaporated under reduced pressure. The crude residue was chromatographed(CHCl₃/AcOEt, 1:1, as eluent) to afford pure 13 as a pale yellow oil(1.32 g, 68% yield). ¹H NMR: δ 1.24 (t, 3H, J=7.1 Hz, CH₂CH₃), 1.79-1.89(m, 2H, CH₂CH₂CO), 2.34 (t, 2H, J=7.3 Hz, COCH₂), 2.41 [t, 2H, J=7.4 Hz,(CH₂)₂NCH₂], 2.63 [br s, 4H, (CH₂)₂NCH₂], 3.06 [br s, 4H, ArN(CH₂)₂],3.83 (s, 3H, OCH₃), 4.11 (q, 2H, J=7.1 Hz, CH₂CH₃), 6.82-7.00 (m, 3H,aromatic). GC-MS m/z 307 (M⁺+1, 18), 306 (M⁺, 77), 261 (32), 205 (100),190 (37).

4-[4-(2-Methoxyphenyl)piperazin-1-yl)]butanoic acid (14)

Ester 13 (1.20 g, 3.9 mmol) was refluxed for 4 h in 20 mL of 4% aqueousNaOH. Then, the mixture was cooled and washed with CHCl₃. The separatedaqueous phase was neutralized with 3 N HCl and extracted with AcOEt (3

30 mL). The collected organic layers were dried over Na₂SO₄ andevaporated under reduced pressure to give 0.58 g of acid 14 as a whitesolid (51% yield). ¹H NMR: δ 1.84-1.89 (m, 2H, CH₂CH₂CO), 2.58-2.62 (m,2H, COCH₂), 2.77 (br t, 2H, (CH₂)₂NCH₂], 2.2.96 [br s, 4H, (CH₂)₂NCH₂],3.20 [br s, 4H, ArN(CH₂)₂], 3.87 (s, 3H, CH₃), 6.87-7.06 (m, 3H,aromatic), 9.52 (br s, 1H, OH, D₂O exchanged). GC-MS m/z 279 (M⁺+1, 20),278 (M⁺, 96), 219 (25), 205 (100), 190 (39).

General Procedure for Preparation of Final Compounds

A stirred mixture of alkylating agent 10a-c,e, 11a-e, 12a-e (8.0 mmol),1-substituted piperazine (9.6 mmol) and K₂CO₃ (8.0 mmol) in acetonitrilewas refluxed overnight. After cooling, the mixture was evaporated todryness and H₂O (20 mL) was added to the residue. The aqueous phase wasextracted with AcOEt (2

30 mL). The collected organic layers were dried over Na₂SO₄ andevaporated under reduced pressure. The crude residue was chromatographed(CH₂Cl₂/CH₃OH, 19:1, as eluent) to yield pure compounds 7, 15-22, 24-43,45-50. as pale yellow oils. Yields were between 20-30% for butanamidederivatives, 35-44% for pentanamide derivatives and 65-75% for the othercompounds.

4-(2-Methoxyphenyl)-N-(1,2,3,4-tetrahydronaphthalen-1-yl)-1-piperazinebutanamide(26)

¹H NMR: δ 1.75-1.93, 1.98-2.10 (m, 6H, COCH₂CH₂, endo CH₂CH₂), 2.34 (t,2H, J=7.0 Hz, COCH₂CH₂), 2.42-2.58 [m, 6H, CH₂N(CH₂)₂], 2.76-2.78 (m,2H, benzylic CH₂), 2.90 [br s, 4H, (CH₂)₂NAr], 3.84 (s, 3H, CH₃),5.19-5.29 (m, 1H, CH), 6.80-7.29 (m, 9H, aromatic, NH). GC-MS m/z 408(M⁺+1, 7), 407 (M⁺, 27), 392 (88), 245 (52), 205 (100).

4-(2-Methoxyphenyl)-N-(1,2,3,4-tetrahydronaphthalen-1-yl)-1-piperazinepentanamide(27)

¹H NMR: δ 1.56-1.85, 2.01-2.07 [m, 8H, CH₂(CH₂)₂CH₂, endo CH₂CH₂], 2.25(t, 2H, J=7.03 Hz, COCH₂CH₂), 2.43 [t, 2H, J=7.3 Hz, CH₂N(CH₂)₂], 2.62[br s, 4H, CH₂N(CH₂)₂], 2.71-2.79 (m, 2H, benzylic CH₂), 3.06 [br s, 4H,(CH₂)₂NAr], 3.86 (s, 3H, CH₃), 5.19-5.23 (m, 1H, CH), 5.79 (br d, 1H,NH), 6.84-7.25 (m, 8H, aromatic). GC-MS m/z 422 (M⁺+1, 4), 421 (M⁺, 14),406 (41), 259 (45), 205 (100), 131 (36).

4-(2-Methoxyphenyl)-N-(1,2,3,4-tetrahydronaphthalen-1-yl)-1-piperazinehexanamide(28)

¹H NMR: δ 1.36-1.43 (m, 2H, CH₂CH₂CH₂CH₂CH₂), 1.51-1.59, 1.61-1.86,2.00-2.06 (m, 8H, CH₂CH₂CH₂CH₂CH₂, endo CH₂CH₂), 2.21 (t, 2H, J=7.6 Hz,COCH₂), 2.40 [br t, 2H, CH₂N(CH₂)₂], 2.64 [br s, 4H, CH₂N(CH₂)₂],2.71-2.80 (m, 2H, benzylic CH₂), 3.09 [br s, 4H, (CH₂)₂NAr], 3.86 (s,3H, CH₃), 5.17-5.23 (m, 1H, CH), 5.67 (br d, 1H, NH), 6.83-7.25 (m, 8H,aromatic). GC-MS m/z 436 (M⁺+1, 4), 435 (M⁺, 13), 420 (27), 273 (41),205 (100).

4-(2-Acetylphenyl)-N-(1,2,3,4-tetrahydronaphthalen-1-yl)-1-piperazinehexanamide(34)

¹H NMR: δ 1.33-1.43 (m, 2H, CH₂CH₂CH₂CH₂CH₂), 1.51-1.86, 1.98-2.06 (m,8H, CH₂CH₂CH₂CH₂CH₂, endo CH₂CH₂), 2.21 (t, 2H, J=7.4 Hz, COCH₂), 2.43[t, 2H, J=7.6 Hz, CH₂N(CH₂)₂], 2.62 [br s, 4H, CH₂N(CH₂)₂], 2.65 (s, 3H,CH₃), 2.71-2.79 (m, 2H, benzylic CH₂), 3.04 [br t, 4H, (CH₂)₂NAr],5.17-5.29 (m, 1H, CH), 5.69 (br d, 1H, NH), 7.02-7.40 (m, 7H, aromatic).GC-MS m/z 448 (M⁺+1, 8), 447 (M⁺, 26), 299 (60), 287 (65), 273 (100),217 (90).

4-(2-Methylthiophenyl)-N-(1,2,3,4-tetrahydronaphthalen-1-yl)-1-piperazinehexanamide(44)

¹H NMR: δ 1.33-1.43 (m, 2H, CH₂CH₂CH₂CH₂CH₂), 1.53-1.63, 1.66-1.86,2.00-2.06 (m, 8H, CH₂CH₂CH₂CH₂CH₂, endo CH₂CH₂), 2.22 (t, 2H, J=7.4 Hz,COCH₂), 2.40 (s, 3H, CH₃), 2.43 [t, 2H, J=7.4 Hz, CH₂N(CH₂)₂], 2.63 [brs, 4H, CH₂N(CH₂)₂], 2.74-2.79 (m, 2H, benzylic CH₂), 3.03 [br s, 4H,(CH₂)₂NAr], 5.18-5.29 (m, 1H, CH), 5.70 (br d, 1H, NH), 7.03-7.26 (m,8H, aromatic). GC-MS m/z 452 (M⁺+1, 2), 451 (M⁺, 8), 273 (61), 221(100).

4-(2-Methylphenyl)-N-(1,2,3,4-tetrahydronaphthalen-1-yl)-1-piperazinehexanamide(49)

¹H NMR: δ 1.34-1.44 (m, 2H, CH₂CH₂CH₂CH₂CH₂), 1.53-1.63, 1.66-1.86,2.00-2.08 (m, 8H, CH₂CH₂CH₂CH₂CH₂, endo CH₂CH₂), 2.19 (t, 2H, J=7.4 Hz,COCH₂), 2.30 (s, 3H, CH₃), 2.42 [br t, 2H, CH₂N(CH₂)₂], 2.60 [br s, 4H,CH₂N(CH₂)₂], 2.70-2.79 (m, 2H, benzylic CH₂), 2.95 [br t, 4H,(CH₂)₂NAr], 5.18-5.23 (m, 1H, CH), 5.69 (br d, 1H, NH), 6.94-7.26 (m,8H, aromatic). GC-MS m/z 420 (M⁺+1, 2), 419 (M⁺, 7), 273 (99), 189(100).

N-(8-Methoxy-1,2,3,4-tetrahydronaphthalen-1-yl)4-(2-methoxyphenyl)-1-piperazinebutanamide(23)

A mixture of carboxylic acid 14 (0.50 g, 1.8 mmol) and1,1′-carbonyldiimidazole (0.29 g, 1.8 mmol) in 10 mL of anhydrous THFwas stirred for 8 h. A solution of amine 9d (0.32 g, 1.8 mmol) in 10 mLof anhydrous THF was added and the resulting mixture was stirred for 1h. The reaction mixture was partitioned between AcOEt and H₂O. Theorganic layer was washed with aqueous Na₂CO₃ solution, dried (Na₂SO₄)and concentrated in vacuo. The crude residue was chromatographed(CH₂Cl₂/CH₃OH, 19:1, as eluent) to afford pure amide 23 (0.33 g, 42%yield). ¹H NMR: δ 1.61-1.90, 2.10-2.19 (m, 6H, COCH₂CH₂, endo CH₂CH₂),2.24 (t, 2H, J=7.4 Hz, COCH₂CH₂), 2.28-2.47, 2.55-2.58 [m, 6H,CH₂N(CH₂)₂], 2.68-2.77 (m, 2H, benzylic CH₂), 2.90 [br s, 4H, (CH₂)₂N],3.79, 3.84 (2 s, 6H, 2 CH₃), 5.27-5.29 (m, 1H, CH), 6.46 (br d, 1H, NH),6.67-7.18 (m, 7H, aromatic). GC-MS m/z 438 (M⁺+1, 1), 437 (M⁺, 4), 422(27), 205 (24), 161 (100).

4-(2-Hydroxyphenyl)-N-(1,2,3,4-tetrahydronaphthalen-1-yl)-1-piperazinehexanamide(46)

A stirred mixture of alkyl bromide 12e (0.36 g, 1.1 mmol) and1-(2-hydroxyphenyl)piperazine (0.29 g, 1.6 mmol) in acetonitrile wasrefluxed overnight. After the mixture was cooled, the solvent wasevaporated in vacuo and a saturated aqueous solution of NaHCO₃ (20 mL)was added to the residue. The aqueous phase was extracted with AcOEt (2

30 mL). The collected organic layers were dried over Na₂SO₄ andevaporated under reduced pressure. The crude residue was chromatographed(CHCl₃/CH₃OH, 19:1, as eluent) to yield pure 46 as a pale yellow oil(0.30 g, 65% yield). ¹H NMR: δ 1.34-1.44 (m, 2H, CH₂CH₂CH₂CH₂CH₂),1.53-1.63, 1.66-1.86, 2.00-2.07 (m, 8H, CH₂CH₂CH₂CH₂CH₂, endo CH₂CH₂),2.22 (t, 2H, J=7.6 Hz, COCH₂), 2.43 [t, 2H, J=7.6 Hz, CH₂N(CH₂)₂], 2.63[br s, 4H, CH₂N(CH₂)₂], 2.75-2.79 (m, 2H, benzylic CH₂), 2.90 [br s, 4H,(CH₂)₂NAr], 5.18-5.29 (m, 1H, CH), 5.69 (br d, 1H, NH), 6.83-7.27 (m,9H, aromatic, OH, 1H D₂O exchanged). GC-MS m/z 422 (M⁺+1, 2), 421 (M⁺,5), 273 (100), 191 (28).

Biological Methods.

General. Male Wistar Hannover rats (200-250 g) and male albinoDunkin-Hartley guinea-pigs (300-350 g) were from Harlan (S. Pietro alNatisone, Italy). The animals were handled according to internationallyaccepted principles for care of laboratory animals (E.E.C. CouncilDirective 86/609, O.J. No. L358, Dec. 18, 1986).

Rat recombinant serotonin 5-HT₇R expressed in HEK-293 cells werepurchased from PerkinElmer-NEN (Betsville, Md., USA).

[³H]-LSD, [³H]-8-OH-DPAT, [³H]-ketanserin were obtained fromPerkinElmer-NEN (Zaventem, Belgium). 5-CT, substance P, and ketanserinwere purchased from Tocris Cookson Ltd. (Bristol, UK). 8-OH-DPAThydrobromide was from RBI. SB-269970 was purchased from Sigma-Aldrich(Milan, Italy).

For receptor binding studies, compounds 5-7, 15-50 were dissolved inabsolute ethanol. For isolated guinea-pig ileum assay, compounds 28, 34,44, 46, 49 were dissolved in Krebs-Henseleit solution, pH 7.4.

Radioligand Binding Assay at Rat Cloned 5-HT₇Rs. Binding of [³H]-LSD atrat cloned 5-HT₇ receptor was performed according to a known method. In1 mL of incubation buffer (50 mM Tris, 10 mM MgCl₂ and 0.5 mM EDTA, pH7.4) were suspended 30 μg of membranes, 2.5 nM [³H]-LSD, the drugs orreference compound (six to nine concentrations). The samples wereincubated for 60 min at 37° C. The incubation was stopped by rapidfiltration on GF/A glass fiber filters (pre-soaked in 0.5%polyethylenimine for 30 min). The filters were washed with 3

3 mL of ice-cold buffer (50 mM Tris, pH 7.4). Nonspecific binding wasdetermined in the presence of 10 μM 5-CT. Approximately 90% of specificbinding was determined under these conditions.

Radioligand Binding Assay at Rat Hippocampal Membranes 5-HT_(1A)Receptors. Binding experiments were performed according to a knownmethod. Rats were killed by decapitation, the brain was quickly removed,and the hippocampus was dissected. The hippocampus (1.0 g) washomogenized with a Brinkman polytron (setting 5 for 3

15 s) in 25 mL of 50 mM Tris buffer, pH 7.6. The homogenate wascentrifuged at 48000 g for 15 min at 4° C. The supernatant wasdiscarded, and the pellet was resuspended in 25 mL of buffer, thenpreincubated for 10 min at 37° C. The homogenate was centrifuged at48000 g for 15 min at 4° C. The supernatant was discarded, and the finalpellet was stored at −80° C. until used. Each tube received in a finalvolume of 1 mL of 50 mM Tris (pH 7.6) hippocampus membranes suspensionand 1 nM [³H]-8-OH-DPAT. For competitive inhibition experiments variousconcentrations of drugs studied were incubated. Nonspecific binding wasdefined using 1 μM 8-OH-DPAT. Samples were incubated at 37° C. for 20min and then filtered on Whatman GF/B glass microfiber filters. TheK_(d) value determined for 8-OH-DPAT was 8.8 nM.

Radioligand Binding Assay at Rat Cortex Membranes 5-HT_(2A) Receptors.Binding experiment was performed according to a known method. Rats werekilled by decapitation, the brain was quickly removed, and the cortexwas dissected. The cortex (1.0 g) was homogenized with a Brinkmanpolytron (setting 5 for 3

15 s) in 25 mL of 0.25 M sucrose. The homogenate was centrifuged at 2000g for 10 min at 4° C. The supernatant was saved, and the pellet wasresuspended in 25 mL of buffer. The surnatantes were collected anddiluted 1:10 w/w with 10 mM Tris pH 7.4. The homogenate wascentrifugated at 35000 g for 15 min at 4° C. The supernatant wasdiscarded, and the final pellet was stored at −80° C. until used. Eachtube received, in a final volume of 2 mL of 50 mM Tris (pH 7.7), cortexmembranes suspension and 2.5 nM [³H]-ketanserin. For competitiveinhibition experiments various concentrations of drugs studied wereincubated. Nonspecific binding was defined using 10 μM ketanserin.Samples were incubated at 37° C. for 15 min and then filtered on WhatmanGF/B glass microfiber filters. The K_(d) value determined for ketanserinwas 0.42 nM.

Isolated Guinea-Pig Ileum Assay. Guinea-pigs were anesthetized and thendecapitated and the proximal ileum removed. The intestine was carefullyflushed several times with warm Krebs-Henseleit solution (118 mM NaCl,25 mM NaHCO₃, 4.7 mM KCl, 0.6 mM MgSO₄, 1.2 mM KH₂PO₄, 1.2 mM CaCl₂,11.2 mM glucose, pH 7.4). Whole ileal segments, of about 3 cm in length,were suspended under 1.0 g tension in Krebs solution gassed with 95% O₂and 5% CO₂ and maintained at 37° C. According to Eglen et al. (supra)with minor modification, the bathing medium contained 1 μM atropine toantagonize cholinergically mediated contractions due to activation of5-HT₃ and 5-HT₄ receptors, 1 μM ketanserin to block 5-HT_(2A) receptors,1 μM pyrilamine to block H₁ receptors. Changes in tension of the tissuewere recorded by Fort 10 Original WPI isometric transducer (2BiologicalInstruments, Italy) connected to a PowerLab/400 workstation. Tissue wascontracted by 100 nM substance P. This value was preliminary determinedby concentration-response curves (1 nM-200 nM). 100 nM substance Pelicited 80% of maximum contraction. The reference agonist 5-CT ortested compound was added 3 min before substance P addition andnon-cumulative concentration-response curves were constructed (0.001μM-10 μM). Because we determined that 5-CT induced relaxation withmaximal response (39%) at 3 μM concentration, 5-HT₇ desensitization wasachieved by equilibrating for 1 h in the presence of 3 μM 5-CT changingthe bathing solution every 15 min. Tested compounds were added 3 minbefore substance P addition.

Full agonists 5-CT, 28, 44, 49 and partial agonist 34 were also testedin the presence of the antagonist SB-269970 (0.1 μM-3 μM). The isolatedguinea pig ileum was equilibrated for 75 min with antagonist beforeconstructing concentration-response curves of tested compounds.

Tissue responses were recorded as gram changes in isometric tension andexpressed as percentage of reduction in the height of the contraction.

Statistical Analysis. The inhibition curves on the different bindingsites of the compounds reported in Table 1 were analyzed by nonlinearcurve fitting utilizing the GRAPHPAD PRISM® program. The value for theinhibition constant, K_(i), was calculated by using the Cheng-Prusoffequation. Agonist potencies, expressed as EC₅₀, were obtained fromnon-linear iterative curve fitting by GRAPHPAD PRISM®.

To the extent that compounds of the general formula I are opticallyactive, the formula I includes both any isolated optical antipodes andthe corresponding optionally racemic mixtures in any conceivablecomposition.

Various embodiments of the invention are foreseen to have valuableapplication as constituents of pharmaceutical preparations to treatvarious conditions generally defined as pathologies. Accordingly,embodiments of the invention also comprise pharmaceutical compositionscomprising one or more compounds of this invention in association with apharmaceutically acceptable carrier. Preferably these compositions arein unit dosage forms such as tablets, pills, capsules, powders,granules, sterile parenteral solutions or suspensions, metered aerosolor liquid sprays, drops, ampoules, auto-injector devices orsuppositories; for oral, parenteral, intranasal, sublingual or rectaladministration, or for administration by inhalation or insufflation. Forpreparing solid compositions such as tablets, the principal activeingredient is mixed with a pharmaceutical carrier, e.g. conventionaltableting ingredients such as corn starch, lactose, sucrose, sorbitol,talc, stearic acid, magnesium stearate, dicalcium phosphate or gums, andother pharmaceutical diluents, e.g. water, to form a solidpreformulation composition containing a homogeneous mixture of acompound of the present invention, or a pharmaceutically acceptable saltthereof. When referring to these preformulation compositions ashomogeneous, it is meant that the active ingredient is dispersed evenlythroughout the composition so that the composition may be readilysubdivided into equally effective unit dosage forms such as tablets,pills and capsules. This solid preformulation composition is thensubdivided into unit dosage forms of the type described above containingfrom 0.1 to about 500 mg of the active ingredient of the presentinvention. Typical unit dosage forms contain from 1 to 100 mg, forexample 1, 2, 5, 10, 25, 50 or 100 mg, of the active ingredient. Thetablets or pills of the novel composition can be coated or otherwisecompounded to provide a dosage form affording the advantage of prolongedaction. For example, the tablet or pill can comprise an inner dosage andan outer dosage component, the latter being in the form of an envelopeover the former. The two components can be separated by an enteric layerwhich serves to resist disintegration in the stomach and permits theinner component to pass intact into the duodenum or to be delayed inrelease. A variety of materials can be used for such enteric layers orcoatings, such materials including a number of polymeric acids andmixtures of polymeric acids with such materials as shellac, cetylalcohol and cellulose acetate.

The liquid forms in which the novel compositions of the presentinvention may be incorporated for administration orally or by injectioninclude aqueous solutions, suitably flavoured syrups, aqueous or oilsuspensions, and flavoured emulsions with edible oils such as cottonseedoil, sesame oil, coconut oil or peanut oil, as well as elixirs andsimilar pharmaceutical vehicles. Suitable dispersing or suspendingagents for aqueous suspensions include synthetic and natural gums suchas tragacanth, acacia, alginate, dextran, sodium carboxymethylcellulose,methylcellulose, polyvinyl-pyrrolidone or gelatin. Thus, based on theabove, a variety of pharmaceutically acceptable doses are provided.

Also, it is noted that the term “pharmaceutically acceptable salt(s)”refers to salts derived from treating a compound of formula 1 with anorganic or inorganic acid such as, for example, acetic, lactic, citric,cinnamic, tartaric, succinic, fumaric, maleic, malonic, mandelic, malic,oxalic, propionic, hydrochloric, hydrobromic, phosphoric, nitric,sulfuric, glycolic, pyruvic, methanesulfonic, ethanesulfonic,toluenesulfonic, salicylic, benzoic, or similarly known acceptableacids.

Toward demonstration of various utilities of embodiments of the presentinvention, the following animal-based example is provided.

Example

Studies with hamsters show that systemic injections of8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT), a serotonergicagonist at the 5-HT_(1A/5A/7) receptors induce circadian phase advances.Microinjection of compound 44 into the dorsal raphe cause phase shiftadvances in a similar manner as did 8-OH-DPAT. Microinjection of theselective 5-HT₇ receptor antagonist SB-26770-A into the dorsal raphebefore microinjection of compound 44 significantly block phase shiftsrelative to pretreatment with vehicle (control). This provides evidencethat no additional receptor subtype, such as the 5-HT_(1A) receptor orthe 5-HT_(5A) receptor, is necessary for this effect.

Based on all of the above teachings and data, it is advanced thateffective treatment of one or more states selected from a circadianrhythm disturbance, a sleep disorder, depression, psychosis, andmigraine is achieved by administering a determined pharmaceuticallyacceptable dosage, comprising one or more of the above-disclosed 5-HT₇receptor agonists or antagonists, to a human or animal subject in needthereof. Such compounds are administered in a variety of forms thatinclude, but are not limited to, pharmaceutical compositions asdescribed above. Most simply, a dosage of one such compounds isadministered to a human in need thereof, for a time regime determinedbased on the particular condition (i.e., pathology) and age, weight,etc. of the subject, and a desired effect with regard to the conditionis obtained based on the selective action of the compound on 5-HT₇receptors.

Various embodiments of the present invention have been shown anddescribed herein, and such embodiments provide teachings of variousaspects of the invention. It is appreciated that variations, changes andsubstitutions may be made beyond these specific disclosures withoutdeparting from the invention herein. Accordingly, it is intended thatthe invention be limited only by the spirit and scope of the appendedclaims in accordance with the relevant law as to their interpretation.

TABLE 1 Physical Properties and Binding Affinities of Compounds 5-7,15-28

K_(i), nM Compd R n formula^(a) mp, ° C. 5-HT₇ 5-HT_(1A)  5^(b) 5-CH₃O 1— — >1000 NT  6^(c) 5-CH₃O 2 — — 269 ± 18  253 ± 25   7 5-CH₃O 3C₂₆H₃₅N₃O₃•HCl•0.2H₂O 215 dec  35 ± 3.2 254 ± 65  15 5-CH₃O 4C₂₇H₃₇N₃O₃•HCl•H₂O 187-190 28.2 ± 4.20 54.4 ± 6.5  16 5-CH₃O 5C₂₈H₃₉N₃O₃•2HCl•0.5H₂O 194-197  20 ± 2.5 24.5 ± 1.8  17 6-CH₃O 3C₂₆H₃₅N₃O₃•2HCl 128-129 186 ± 40  257 ± 25  18 6-CH₃O 4 C₂₇H₃₇N₃O₃•2HCl149-152 43.1 ± 4.8  23.2 ± 2.3  19 6-CH₃O 5 C₂₈H₃₉N₃O₃•(COOH)₂ 102-105  30 ± 3.15  55 ± 8.0 20 7-CH₃O 3 C₂₆H₃₅N₃O₃•2HCl•H₂O 118-120 129 ± 5.0 160 ± 12  21 7-CH₃O 4 C₂₇H₃₇N₃O₃•2HCl•0.4H₂O 187-188 38.4 ± 4.6  78.9 ±6.30 22 7-CH₃O 5 C₂₈H₃₉N₃O₃•2HCl 175-178 41 ± 11  39 ± 6.5 23 8-CH₃O 3C₂₆H₃₅N₃O₃•2HCl 131-133 154 ± 35  441 ± 20  24 8-CH₃O 4C₂₇H₃₇N₃O₃•2HCl•0.2H₂O 136-137 64.0 ± 12   72.0 ± 18   25 8-CH₃O 5C₂₈H₃₉N₃O₃•2HCl 125-127 31.4 ± 3.5  30.0 ± 2.6  26 H 3 C₂₅H₃₃N₃O₂•2HCl171-173 92.0 ± 12   245 ± 20  27 H 4 C₂₆H₃₅N₃O₂•2HCl•0.8H₂O 199-200 6.05± 0.25   9 ± 0.70 28 H 5 C₂₇H₃₇N₃O₂•2HCl•0.5H₂O 150-151 6.64 ± 0.60  8.6± 0.35 5-CT 0.51 ± 0.01 8-OH-DPAT 1.2 ± 0.2 ^(a)All compounds wererecrystallized from CH₃OH/Et₂O. Analysis for C, H, N; results werewithin ±0.4% of the theoretical values for the formulas given. ^(b)Seeref. 33. ^(c)See ref. 32.

TABLE 2 Physical Properties and Binding Affinities of Compounds 29-32

K_(i), nM compd Ar formula^(a) mp, ° C. 5-HT₇ 5-HT_(1A) 29

C₂₇H₃₄N₄O₂•2HCl•H₂O 164 dec 125 ± 30 3900 ± 120 30

C₂₇H₃₄N₄O₂ 148 dec 820 ± 90 NT^(b) 31

C₂₇H₃₅N₅O•3HCl 300 dec 356 ± 55 2600 ± 280 32

C₂₈H₃₆N₄O₃•2HCl•H₂O 196-198 704 ± 30 NT ^(a)All compounds wererecrystallized from CH₃OH/Et₂O except 30 (from CHCl₃/n-hexane). Analysisfor C, H, N; results were within ±0.4% of the theoretical values for theformulas given. ^(b)Not tested.

TABLE 3 Physical Properties and Binding Affinities of Compounds 33-50

K_(i), nM cpd R formula^(a) mp, ° C. 5-HT₇ 5-HT_(1A) 5-HT_(2A) 33 2-CNC₂₇H₃₄N₄O•HCl•0.5H₂O 175-178 48.7 ± 2.5  16.6 ± 1.4  700 ± 25  342-COCH₃ C₂₈H₃₇N₃O₂•HCl•H₂O 136-139 4.14 ± 0.80  3.8 ± 0.10 12200 ± 350 35 2-OCH₃-4-Cl C₂₇H₃₆N₃O₂Cl 145-147 122 ± 14  332 ± 27  7168 ± 150  362,5-di-OCH₃ C₂₈H₃₉N₃O₃•2HCl•0.6H₂O 124-126 70.3 ± 5.2  911 ± 23  259 ±20  37 3-OCH₃ C₂₇H₃₇N₃O₂•2HCl•0.5H₂O 156-159 119 ± 20  105 ± 12  142 ±20  38 4-OCH₃ C₂₇H₃₇N₃O₂ 129-130 2100 ± 150  NT^(b) NT 39 3-CNC₂₇H₃₄N₄O•HCl 170-172 97.8 ± 5.6  291 ± 15  909 ± 85  40 4-CNC₂₇H₃₄N₄O•HCl•1.5H₂O 102-104 1400 ± 120  NT NT 41 3-COCH₃C₂₈H₃₇N₃O₂•2HCl•H₂O 146-148 496 ± 24  676 ± 32  1127 ± 200  42 4-COCH₃C₂₈H₃₇N₃O₂•2HCl 112-114 2639 ± 130  NT NT 43 2-CONH₂C₂₇H₃₆N₄O₂•2HCl•0.5H₂O 184-187 229 ± 12  494 ± 35  >4000 (9%)  44 2-SCH₃C₂₇H₃₇N₃OS•HCl•H₂O 181-182 0.22 ± 0.08 52.7 ± 3.2  326 ± 35  45 2-SO₂CH₃C₂₇H₃₇N₃O₃S•HCl•H₂O 120-122 298 ± 16  3124 ±260  >4000 (38%) 46 2-OHC₂₆H₃₅N₃O₂•2HCl•0.3H₂O 162-164 11.4 ± 2.3   24 ± 6.3 3394 ± 225  47 2-ClC₂₆H₃₄ClN₃O•HCl•0.3H₂O 168-169 40.1 ± 6.7   96 ± 8.0 301 ± 12  48 2-NO₂C₂₆H₃₄N₄O₃•HCl•0.5H₂O 152-155 63.3 ± 7.5  183 ± 15  282 ± 35  49 2-CH₃C₂₇H₃₇N₃O•2HCl•0.5H₂O 212-214 15.2 ± 3.2  279 ± 44  262 ± 24  50 HC₂₆H₃₅N₃O•2HCl•0.5H₂O 172-174 65.6 ± 4.7  128 ± 22  77.8 ± 5.7  ^(a)Allcompounds were recrystallized from CH₃OH/Et₂O except 35 and 38 (fromCHCl₃/n-hexane). Analysis for C, H, N; results were within ±0.4% of thetheoretical values for the formulas given. ^(b)Not tested.

TABLE 4 Relaxation effect induced by selected compounds and 5-CT onsubstance P-stimulated guinea pig ileum contracture with 5-HT₇ receptordesensitization. Maximal PA₂ cpd Effect % EC₅₀, μM (vs SB-269970) Schildplot N 28 91 6.32 ± 0.20 8.02 ± 1.40 1.0 (p < 0.0001) 15 34 79 2.46 ±0.70 7.60 ± 0.49 1.3 (p < 0.0001) 14 44 100 2.56 ± 0.32 7.70 ± 0.80 0.9(p < 0.0001) 15 46 0 — 7.20 ± 0.60 1.6 (p < 0.005)  16 49 98 1.82 ± 0.727.80 ± 0.40 0.9 (p < 0.0001) 15 5-CT 100 0.63 ± 0.04 7.48 ± 0.12 1.2 (p< 0.0001) 12

1. 5-hydroxytryptamine-7 (5-HT₇) receptor ligands, having generalformula I

wherein: R means 2-methoxy, 2-acetyl, 2-methylthio, 2-hydroxy, 2-methylgroup and n means 4 or 5 or pharmaceutically acceptable salts thereof.2. The 5-HT₇ receptor ligands of claim 1, wherein n is 5 and R is amethylthio group or n is 5 and R is a hydroxy group or n is 5 and R is amethyl group or n is 5 and R is a methoxy group or n is 4 and R is amethoxy group.
 3. 5-HT₇ receptor ligands according to claim 1, wherein nis 5 and R is a methylthio group.
 4. 5-HT₇ receptor ligands according toclaim 1, wherein n is 5 and R is a hydroxy group.
 5. 5-HT₇ receptorligands according to claim 1, wherein n is 5 and R is a methyl group. 6.5-HT₇ receptor ligands according to claim 1, wherein n is 5 and R is amethoxy group.
 7. 5-HT₇ receptor ligands according to claim 1, wherein nis 4 and R is a methoxy group.
 8. Pharmaceutic composition comprising atherapeutically effective amount of one or more 5-HT₇ receptor ligandsaccording to claim 1 or a pharmaceutically acceptable salt thereof and apharmaceutically acceptable excipient.
 9. Pharmaceutic compositionaccording to claim 8, comprising one or more ligands or pharmaceuticallyacceptable salts thereof selected from the group consisting of4-(2-Methylthiophenyl)-N-(1,2,3,4-tetrahydronaphthalen-1-yl)-1-piperazinehexanamide(44);4-(2-Methylphenyl)-N-(1,2,3,4-tetrahydronaphthalen-1-yl)-1-piperazinehexanamide(49);4-(2-Methoxyphenyl)-N-(1,2,3,4-tetrahydronaphthalen-1-yl)-1-piperazinehexanamide(28);4-(2-Methoxyphenyl)-N-(1,2,3,4-tetrahydronaphthalen-1-yl)-1-piperazinepentanamide(27);4-(2-Acetylphenyl)-N-(1,2,3,4-tetrahydronaphthalen-1-yl)-1-piperazinehexanamide(34);4-(2-Hydroxyphenyl)-N-(1,2,3,4-tetrahydronaphthalen-1-yl)-1-piperazinehexanamide(46) and a pharmaceutically acceptable excipient.
 10. A method oftreatment pathological states suitable to be relieved by 5-HT₇ receptoragonists or antagonists comprising the step of administering one or more5-hydroxytryptamine-7 (5-HT₇) receptor ligands according to claim 1 to asubject in need, wherein the pathological state is selected from thegroup consisting of circadian rhythm disturbances and depression.
 11. Amethod of preparing a compound according to claim 1 comprising acylatingamines of formula 9e:

with the appropriate ω-haloacyl chloride of the following formula:

to obtain the corresponding intermediate compounds of formula 11ewherein X is chlorine or 12e wherein X is bromine:

reacting these intermediate compounds with the appropriate1-(2-substituted phenyl) piperazine of the following formula:

and optionally reacting the obtained compound of formula I with asuitable acid to prepare its pharmaceutically acceptable salt, wherein Rmeans 2-methoxy, 2-acetyl, 2-methylthio, 2-hydroxy, of 2-methyl group,and n means 4 or
 5. 12. The method of claim 11, additionally comprisingreacting the compound of Formula 1 with a suitable acid to prepare itspharmaceutically acceptable salt.